Abstract
Key message
DcWRKY5 increases the antioxidant enzyme activity and proline accumulation, oppositely, reduces the accumulation of ROS and MDA, through directly activating the genes expression, finally enhances the salt and drought tolerance.
Abstract
Drought and salinity are two main environmental factors that limit the large-scale cultivation of the medicinal plant Dioscorea composita (D. composita). WRKY transcription factors (TFs) play vital roles in regulating drought and salt tolerance in plants. Nevertheless, the molecular mechanism of WRKY TF mediates drought and salt resistance of D. composita remains largely unknown. Here, we isolated and characterized a WRKY TF from D. composita, namely DcWRKY5, which was localized to the nucleus and bound to the W-box cis-acting elements. Expression pattern analysis showed that it was highly expressed in root and significantly up-regulated in the presence of salt, polyethylene glycol-6000 (PEG-6000) and abscisic acid (ABA). Heterologous expression of DcWRKY5 increased salt and drought tolerance in Arabidopsis, but was insensitive to ABA. In addition, compared with the wild type, the DcWRKY5 overexpressing transgenic lines had more proline, higher antioxidant enzyme (POD, SOD, and CAT) activities, less reactive oxygen species (ROS) and malondialdehyde (MDA). Correspondingly, the overexpression of DcWRKY5 modulated the expression of genes related to salt and drought stresses, such as AtSS1, AtP5CS1, AtCAT, AtSOD1, AtRD22, and AtABF2. Dual luciferase assay and Y1H were further confirmed that DcWRKY5 activate the promoter of AtSOD1 and AtABF2 through directly binding to the enrichment region of the W-box cis-acting elements. These results suggest that DcWRKY5 is a positive regulator of the drought and salt tolerance in D. composita and has potential applications in transgenic breeding.
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References
Bai Y, Sunarti S, Kissoudis C, Visser RGF, van der Linden CG (2018) The role of tomato WRKY genes in plant responses to combined abiotic and biotic stresses. Front Plant Sci 9:801. https://doi.org/10.3389/fpls.2018.00801
Bao W, Wang X, Chen M, Chai T, Wang H (2018) A WRKY transcription factor, PcWRKY33, from Polygonum cuspidatum reduces salt tolerance in transgenic Arabidopsis thaliana. Plant Cell Rep 37(7):1033–1048. https://doi.org/10.1007/s00299-018-2289-2
Bo C, Cai R, Fang X, Wu H, Ma Z, Yuan H, Cheng B, Fan J, Ma Q (2022) Transcription factor ZmWRKY20 interacts with ZmWRKY115 to repress expression of ZmbZIP111 for salt tolerance in maize. Plant J. https://doi.org/10.1111/tpj.15914
Chen X, Li C, Wang H, Guo Z (2019) WRKY transcription factors: evolution, binding, and action. Phytopathol Res. https://doi.org/10.1186/s42483-019-0022-x
Chu X, Wang C, Chen X, Lu W, Li H, Wang X, Hao L, Guo X (2015) The cotton WRKY gene GhWRKY41 positively regulates salt and drought stress tolerance in transgenic Nicotiana benthamiana. PLoS ONE 10(11):e0143022. https://doi.org/10.1371/journal.pone.0143022
Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16(6):735–743. https://doi.org/10.1046/j.1365-313x.1998.00343.x
Deinlein U, Stephan AB, Horie T, Luo W, Xu G, Schroeder JI (2014) Plant salt-tolerance mechanisms. Trends Plant Sci 19(6):371–379. https://doi.org/10.1016/j.tplants.2014.02.001
Dietz KJ, Zorb C, Geilfus CM (2021) Drought and Crop Yield. Plant Biol (Stuttg) 23(6):881–893. https://doi.org/10.1111/plb.13304
Dong Q, Zheng W, Duan D, Huang D, Wang Q, Liu C, Li C, Gong X, Li C, Mao K, Ma F (2020) MdWRKY30, a group IIa WRKY gene from apple, confers tolerance to salinity and osmotic stresses in transgenic apple callus and Arabidopsis seedlings. Plant Sci 299:110611. https://doi.org/10.1016/j.plantsci.2020.110611
Erpen L, Devi HS, Grosser JW, Dutt M (2017) Potential use of the DREB/ERF, MYB, NAC and WRKY transcription factors to improve abiotic and biotic stress in transgenic plants. Plant Cell Tiss Organ Cult (PCTOC) 132(1):1–25. https://doi.org/10.1007/s11240-017-1320-6
Gao YF, Liu JK, Yang FM, Zhang GY, Wang D, Zhang L, Ou YB, Yao YA (2020) The WRKY transcription factor WRKY8 promotes resistance to pathogen infection and mediates drought and salt stress tolerance in Solanum lycopersicum. Physiol Plant 168(1):98–117. https://doi.org/10.1111/ppl.12978
Goyal P, Devi R, Verma B, Hussain S, Arora P, Tabassum R, Gupta S (2022) WRKY transcription factors: evolution, regulation, and functional diversity in plants. Protoplasma. https://doi.org/10.1007/s00709-022-01794-7
Han G, Lu C, Guo J, Qiao Z, Sui N, Qiu N, Wang B (2020) C2H2 zinc finger proteins: master regulators of abiotic stress responses in plants. Front Plant Sci 11:115. https://doi.org/10.3389/fpls.2020.00115
Huang Y, Feng CZ, Ye Q, Wu WH, Chen YF (2016) Arabidopsis WRKY6 transcription factor acts as a positive regulator of abscisic acid signaling during seed germination and early seedling development. PLoS Genet 12(2):e1005833. https://doi.org/10.1371/journal.pgen.1005833
Jiang J, Ma S, Ye N, Jiang M, Cao J, Zhang J (2017) WRKY transcription factors in plant responses to stresses. J Integr Plant Biol 59(2):86–101. https://doi.org/10.1111/jipb.12513
Jiang Y, Tong S, Chen N, Liu B, Bai Q, Chen Y, Bi H, Zhang Z, Lou S, Tang H, Liu J, Ma T, Liu H (2021) The PalWRKY77 transcription factor negatively regulates salt tolerance and abscisic acid signaling in Populus. Plant J 105(5):1258–1273. https://doi.org/10.1111/tpj.15109
Ju YL, Yue XF, Min Z, Wang XH, Fang YL, Zhang JX (2020) VvNAC17, a novel stress-responsive grapevine (Vitis vinifera L.) NAC transcription factor, increases sensitivity to abscisic acid and enhances salinity, freezing, and drought tolerance in transgenic Arabidopsis. Plant Physiol Biochem 146:98–111. https://doi.org/10.1016/j.plaphy.2019.11.002
Kang G, Yan D, Chen X, Yang L, Zeng R (2021) HbWRKY82, a novel IIc WRKY transcription factor from Hevea brasiliensis associated with abiotic stress tolerance and leaf senescence in Arabidopsis. Physiol Plant 171(1):151–160. https://doi.org/10.1111/ppl.13238
Kidokoro S, Watanabe K, Ohori T, Moriwaki T, Maruyama K, Mizoi J, Htwe NMPS, Fujita Y, Sekita S, Shinozaki K, Yamaguchi-Shinozaki K (2015) Soybean DREB1/CBF-type transcription factors function in heat and drought as well as cold stress-responsive gene expression. Plant J 81(3):505–518. https://doi.org/10.1111/tpj.12746
Kumar S, Stecher G, Tamura K (2016) Molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol. 33(7):1870–1874. https://doi.org/10.1093/molbev/msw054
Lee FC, Yeap WC, Appleton DR, Ho CL, Kulaveerasingam H (2022) Identification of drought responsive Elaeis guineensis WRKY transcription factors with sensitivity to other abiotic stresses and hormone treatments. BMC Genom 23(1):164. https://doi.org/10.1186/s12864-022-08378-y
Li JB, Luan YS, Liu Z (2015) Overexpression of SpWRKY1 promotes resistance to Phytophthora nicotianae and tolerance to salt and drought stress in transgenic tobacco. Physiol Plant 155(3):248–266. https://doi.org/10.1111/ppl.12315
Li W, Pang S, Lu Z, Jin B (2020) Function and mechanism of WRKY transcription factors in abiotic stress responses of plants. Plants (basel) 9(11):1515. https://doi.org/10.3390/plants9111515
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2–△△CT method. Methods 25(4):402–408. https://doi.org/10.1006/meth.2001.1262
Luo X, Bai X, Sun X, Zhu D, Liu B, Ji W, Cai H, Cao L, Wu J, Hu M, Liu X, Tang L, Zhu Y (2013) Expression of wild soybean WRKY20 in Arabidopsis enhances drought tolerance and regulates ABA signalling. J Exp Bot 64(8):2155–2169. https://doi.org/10.1093/jxb/ert073
Ma Q, Xia Z, Cai Z, Li L, Cheng Y, Liu J, Nian H (2018) GmWRKY16 enhances drought and salt tolerance through an ABA-mediated pathway in Arabidopsis thaliana. Front Plant Sci 9:1979. https://doi.org/10.3389/fpls.2018.01979
Machens F, Becker M, Umrath F, Hehl R (2014) Identification of a novel type of WRKY transcription factor binding site in elicitor-responsive cis-sequences from Arabidopsis thaliana. Plant Mol Biol 84(4–5):371–385. https://doi.org/10.1007/s11103-013-0136-y
Manna M, Thakur T, Chirom O, Mandlik R, Deshmukh R, Salvi P (2020) Transcription factors as key molecular target to strengthen the drought stress tolerance in plants. Physiol Plant. https://doi.org/10.1111/ppl.13268
Phukan UJ, Jeena GS, Shukla RK (2016) WRKY transcription factors: molecular regulation and stress responses in plants. Front Plant Sci 7:760. https://doi.org/10.3389/fpls.2016.00760
Rajappa S, Krishnamurthy P, Kumar PP (2020) Regulation of AtKUP2 expression by bHLH and WRKY transcription factors helps to confer increased salt tolerance to Arabidopsis thaliana plants. Front Plant Sci 11:1311. https://doi.org/10.3389/fpls.2020.01311
Raza A, Razzaq A, Mehmood SS, Zou X, Zhang X, Lv Y, Xu J (2019) Impact of climate change on crops adaptation and strategies to tackle its outcome: a review. Plants (basel). https://doi.org/10.3390/plants8020034
Rushton PJ, Somssich IE, Ringler P, Shen QJ (2010) WRKY transcription factors. Trends Plant Sci 15(5):247–258. https://doi.org/10.1016/j.tplants.2010.02.006
Ryu H, Cho Y-G (2015) Plant hormones in salt stress tolerance. J Plant Biol 58(3):147–155. https://doi.org/10.1007/s12374-015-0103-z
Sachdev S, Ansari SA, Ansari MI, Fujita M, Hasanuzzaman M (2021) Abiotic stress and reactive oxygen species: generation, signaling, and defense mechanisms. Antioxidants (basel) 10(2):277. https://doi.org/10.3390/antiox10020277
Sharma P, Jha AB, Dubey RS, Pessarakli M (2012) Reactive oxygen species, oxidative damage, and antioxidative defense mechanism in plants under stressful conditions. J Bot 2012:1–26. https://doi.org/10.1155/2012/217037
Sharma A, Kumar V, Shahzad B, Ramakrishnan M, Singh Sidhu GP, Bali AS, Handa N, Kapoor D, Yadav P, Khanna K, Bakshi P, Rehman A, Kohli SK, Khan EA, Parihar RD, Yuan H, Thukral AK, Bhardwaj R, Zheng B (2019) Photosynthetic response of plants under different abiotic stresses: a review. J Plant Growth Regul 39(2):509–531. https://doi.org/10.1007/s00344-019-10018-x
Shinozaki K, Yamaguchi-Shinozaki K (2007) Gene networks involved in drought stress response and tolerance. J Exp Bot 58(2):221–227. https://doi.org/10.1093/jxb/erl164
Song J, Sun P, Kong W, Xie Z, Li C, Liu JH (2023) SnRK2.4-mediated phosphorylation of ABF2 regulates ARGININE DECARBOXYLASE expression and putrescine accumulation under drought stress. New Phytol 238(1):216–236. https://doi.org/10.1111/nph.18526
Sun TT, Wang C, Liu R, Zhang Y, Wang YC, Wang LQ (2021) ThHSFA1 confers salt stress tolerance through modulation of reactive oxygen species scavenging by directly regulating ThWRKY4. Int J Mol Sci 22(9):5048. https://doi.org/10.3390/ijms22095048
Takahashi F, Shinozaki K (2019) Long-distance signaling in plant stress response. Curr Opin Plant Biol 47:106–111. https://doi.org/10.1016/j.pbi.2018.10.006
Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30(12):2725–2729. https://doi.org/10.1093/molbev/mst197
Tomar RS, Kataria S, Jajoo A (2021) Behind the scene: critical role of reactive oxygen species and reactive nitrogen species in salt stress tolerance. J Agron Crop Sci 207(4):577–588. https://doi.org/10.1111/jac.12490
Ulker B, Somssich IE (2004) WRKY transcription factors: from DNA binding towards biological function. Curr Opin Plant Biol 7(5):491–498. https://doi.org/10.1016/j.pbi.2004.07.012
Ullah A, Sun H, Hakim YX, Zhang X (2018) A novel cotton WRKY gene, GhWRKY6-like, improves salt tolerance by activating the ABA signaling pathway and scavenging of reactive oxygen species. Physiol Plant 162(4):439–454. https://doi.org/10.1111/ppl.12651
van Zelm E, Zhang Y, Testerink C (2020) Salt tolerance mechanisms of plants. Annu Rev Plant Biol 71:403–433. https://doi.org/10.1146/annurev-arplant-050718-100005
Wang X, Chen D, Wang Y, Xie J (2015) De novo transcriptome assembly and the putative biosynthetic pathway of steroidal sapogenins of Dioscorea composita. PLoS ONE 10(4):e0124560. https://doi.org/10.1371/journal.pone.0124560
Wang L, Yu G, Macho AP, Lozano-Duran R (2021) Split-luciferase complementation imaging assay to study protein-protein interactions in Nicotiana benthamiana. Bio Protoc 11(23):e4237. https://doi.org/10.21769/BioProtoc.4237
Wani SH, Anand S, Singh B, Bohra A, Joshi R (2021) WRKY transcription factors and plant defense responses: latest discoveries and future prospects. Plant Cell Rep 40(7):1071–1085. https://doi.org/10.1007/s00299-021-02691-8
Willems E, Leyns L, Vandesompele J (2008) Standardization of real-time PCR gene expression data from independent biological replicates. Anal Biochem 379(1):127–129. https://doi.org/10.1016/j.ab.2008.04.036
Wu X, Shiroto Y, Kishitani S, Ito Y, Toriyama K (2009) Enhanced heat and drought tolerance in transgenic rice seedlings overexpressing OsWRKY11 under the control of HSP101 promoter. Plant Cell Rep 28(1):21–30. https://doi.org/10.1007/s00299-008-0614-x
Xiong C, Zhao S, Yu X, Sun Y, Li H, Ruan C, Li J (2020) Yellowhorn drought-induced transcription factor XsWRKY20 acts as a positive regulator in drought stress through ROS homeostasis and ABA signaling pathway. Plant Physiol Biochem 155:187–195. https://doi.org/10.1016/j.plaphy.2020.06.037
Yan J, Li J, Zhang H, Liu Y, Zhang A (2022) ZmWRKY104 positively regulates salt tolerance by modulating ZmSOD4 expression in maize. Crop J 10(2):555–564. https://doi.org/10.1016/j.cj.2021.05.010
Yang Y, Bocs S, Fan H, Armero A, Baudouin L, Xu P, Xu J, This D, Hamelin C, Iqbal A, Qadri R, Zhou L, Li J, Wu Y, Ma Z, Issali AE, Rivallan R, Liu N, Xia W, Peng M, Xiao Y (2021) Coconut genome assembly enables evolutionary analysis of palms and highlights signaling pathways involved in salt tolerance. Commun Biol 4(1):105. https://doi.org/10.1038/s42003-020-01593-x
Yu S, Lan X, Zhou J, Gao K, Zhong C, Xie J (2022) Dioscorea composita WRKY3 positively regulates salt-stress tolerance in transgenic Arabidopsis thaliana. J Plant Physiol 269:153592. https://doi.org/10.1016/j.jplph.2021.153592
Zhang X, Chen L, Shi Q, Ren Z (2020a) SlMYB102, an R2R3-type MYB gene, confers salt tolerance in transgenic tomato. Plant Sci 291:110356. https://doi.org/10.1016/j.plantsci.2019.110356
Zhang Y, Zhou Y, Zhang D, Tang X, Li Z, Shen C, Han X, Deng W, Yin W, Xia X (2020b) PtrWRKY75 overexpression reduces stomatal aperture and improves drought tolerance by salicylic acid-induced reactive oxygen species accumulation in poplar. Environ Exp Bot. https://doi.org/10.1016/j.envexpbot.2020.104117
Zhang T, Li Y, Kang Y, Wang P, Li W, Yu W, Wang J, Wang J, Song X, Jiang X, Zhou Y (2022) The Dendrobium catenatum DcCIPK24 increases drought and salt tolerance of transgenic Arabidopsis. Ind Crops Prod. https://doi.org/10.1016/j.indcrop.2022.115375
Zheng J, Zhang Z, Tong T, Fang Y, Zhang X, Niu C, Li J, Wu Y, Xue D, Zhang X (2021) Genome-wide identification of WRKY gene family and expression analysis under abiotic stress in Barley. Agronomy 11(3):521. https://doi.org/10.3390/agronomy11030521
Zhong C, Tang Y, Pang B, Li X, Yang Y, Deng J, Feng C, Li L, Ren G, Wang Y, Peng J, Sun S, Liang S, Wang X (2020) The R2R3-MYB transcription factor GhMYB1a regulates flavonol and anthocyanin accumulation in Gerbera hybrida. Hortic Res 7:78. https://doi.org/10.1038/s41438-020-0296-2
Zhu JK (2016) Abiotic stress signaling and responses in plants. Cell 167(2):313–324. https://doi.org/10.1016/j.cell.2016.08.029
Zhu D, Hou L, Xiao P, Guo Y, Deyholos MK, Liu X (2019) VvWRKY30, a grape WRKY transcription factor, plays a positive regulatory role under salinity stress. Plant Sci 280:132–142. https://doi.org/10.1016/j.plantsci.2018.03.018
Zorb C, Geilfus CM, Dietz KJ (2019) Salinity and crop yield. Plant Biol (Stuttg) 21(Suppl 1):31–38. https://doi.org/10.1111/plb.12884
Acknowledgements
We acknowledge the funding support from the National Key Research and Development Program of China (2019YFB1503805 and 2019YFB1503802), the National Natural Science Foundation of China (31700282), and the Natural Science Foundation of Guangdong Province (2021A1515011315). We thank 'bio-brushes' for the illustrations of Arabidopsis thaliana. We thank Prof. Shaohua Zeng from South China Botanical Garden for kindly providing the dual-LUC plasmid.
Funding
National Key Research and Development Program of China, 2019YFB1503805, Jun Xie, 2019YFB1503802, Jun Xie, National Natural Science Foundation of China, 31700282, Zhong Chunmei, Natural Science Foundation of Guangdong Province, 2021A1515011315, Zhong Chunmei.
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JX, CZ, and SY jointly conceived and designed the experiment. SY, LY, KG, JZ and XL completed the experiments. SY conducted data analysis and wrote the manuscript. JX and CZ proofread the manuscript. All authors have read and approved the final manuscript.
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Yu, S., Yang, L., Gao, K. et al. Dioscorea composita WRKY5 positively regulates AtSOD1 and AtABF2 to enhance drought and salt tolerances. Plant Cell Rep 42, 1365–1378 (2023). https://doi.org/10.1007/s00299-023-03038-1
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DOI: https://doi.org/10.1007/s00299-023-03038-1